MOLECULAR DESCRIPTION OF THE CORTEX AND HIPPOCAMPUS

我们已经发展了消减杂交技术， 一种分离mRNA克隆的有力工具， 解剖学定义的神经元子集。 利用这项技术， 分离RDS神经元变性基因的产物。 我们提出 用于测试其作为管腔内粘连的功能的模型的研究 分子，以提高我们对rds分布的理解 外节盘膜中的蛋白质，以证明失活 的rds蛋白是唯一负责的rds表型，并 充分了解rds致突变事件和突变体的性质 蛋白质表达 我们还分离了编码A的mRNA克隆 端脑富集、钙调蛋白结合蛋白激酶底物 C和端脑富集Ca++、钙调素依赖蛋白 激酶。 为了解释每一个的生理意义，我们将 绘制它们的基因图谱，以了解它们是否是 已知基因的突变体已被表征，我们也将 产生影响其表达的故意的小鼠突变体。 本 最后，我们将开发核酶和同源重组基因， 敲除方法。 我们将生产重组蛋白， 功能的生化评估，包括证明激酶 真正具有这种活性并寻找其天然基质。 我们 将研究指导蛋白质树突的氨基酸序列。

We have developed the technique of subtractive hybridization as a powerful tool for isolating clones of mRNAs that are restricted to anatomically-defined subsets of neurons. Using this technique we have isolated the product of the rds neuronal degeneration gene. We propose studies to test a model for its function as an intralumenal adhesion molecule, to improve our understanding of the disposition of the rds protein in the outer segment disc membrane, to prove that inactivation of the rds protein is solely responsible for the rds phenotype, and to understand fully the nature of the rds mutagenic event and mutant protein expression. We have also isolated clones of mRNAs encoding a telencephalon-enriched, calmodulin-binding substrate for protein kinase C and a telencephalon-enriched Ca++, calmodulin-dependent protein kinase. To interpret the physiological significance of each, we will map their genes so as to learn whether they are the products of any known gene for which mutants have been characterized and we will also produce deliberate mouse mutants affected in their expression. To this end, we will develop ribozyme and homologous recombination gene knock-out methodologies. We will produce recombinant proteins for biochemical assessments of function, including proof that the kinase truly has that activity and a search for its natural substrates. We will investigate amino acid sequences that direct proteins to dendrites.